Preferential expression and immunogenicity of HIV-1 Tat fusion protein expressed in tomato plant |
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| Authors: | Marni E. Cueno Yurina Hibi Katsuo Karamatsu Yasuhiro Yasutomi Kenichi Imai Antonio C. Laurena Takashi Okamoto |
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| Affiliation: | 1.Department of Molecular and Cellular Biology? Molecular and Cellular Biology Laboratory, Graduate School of Medical Sciences,Nagoya City University,Nagoya,Japan;2.Laboratory of Immunoregulation and Vaccine Research, Tsukuba Primate Research Center,National Institute of Biomedical Innovation,Tsukuba,Japan;3.Biochemistry Laboratory, Institute of Plant Breeding,University of the Philippines Los Banos,Laguna,Philippines |
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| Abstract: | HIV-1 Tat plays a major role in viral replication and is essential for AIDS development making it an ideal vaccine target providing that both humoral and cellular immune responses are induced. Plant-based antigen production, due to its cheaper cost, appears ideal for vaccine production. In this study, we created a plant-optimized tat and mutant (Cys30Ala/Lys41Ala) tat (mtat) gene and ligated each into a pBI121 expression vector with a stop codon and a gusA gene positioned immediately downstream. The vector construct was bombarded into tomato leaf calli and allowed to develop. We thus generated recombinant tomato plants preferentially expressing a Tat-GUS fusion protein over a Tat-only protein. In addition, plants bombarded with either tat or mtat genes showed no phenotypic difference and produced 2–4 μg Tat-GUS fusion protein per milligram soluble plant protein. Furthermore, tomato extracts intradermally inoculated into mice were found to induce a humoral and, most importantly, cellular immunity. |
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